Gene Imaging for Studying Pharmacokinetics of Gene in Vivo at the Real Time

基因成像用于实时研究基因体内药代动力学

• 批准号: 16591225
• 负责人: NAKAMURA Kayoko
• 金额: $ 2.24万
• 依托单位: Keio University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16591225/
• 关键词: Tc-99m; gene imaging; antisense; gene; oligonucleotide; multi drug resistance; mdr1; transfecting reagents

The aim of this study was to develop the gene imaging for studying where the mRNA is located, how much it overexpress, and how it is distributed by using the nuclear medicine modality. Human epidermoid carcinoma cell line; KB-31, its mdr1 transfected cell line; KB-G2, and human thyroid tumor; TCO-1 which overexpress mdr1, were used throughout the in vitro and in vivo studies.1. Antisense DNA (AS) and sense DNA (S) were labeled with Tc-99m under supervised by Dr. Donald J Hnatowich (Professor, University Massachusetts Medical School), and then they were incubated with KB-31, KB-G2 and TCO-1 cells. We found AS-accumulations occurred by the antisense mechanism.2. Gene imaging was successfully obtained when Tc-99m-AS was injected into the KB-G2 tumor implanted into the thigh of mice. This is the first report of antisense imaging in the world.3. Tc-99m-AS was accumulated specifically into the KB-G2 tumors when it was injected into mice intravenously, but its accumulation was not high enough to get the image.4. Transfecting agents such as jetPEI, Neophectin and Chariot enhanced the accumulation of Tc-99m-AS into KB-G2 cells with 3-8-fold comparing with the naked AS-DNAs, but these increases were not observed significantly in vivo study.5. Nanoparticles consisting of Tc-99m-Streptavidine-TAT-AS were prepared by our unique methods and these showed the higher accumulations into the cell comparing with naked AS-DNAs. In vivo studies are now undergoing.

本研究的目的是通过核医学模式开发用于研究mRNA定位的基因成像，其过度表达的程度以及其如何分布。本研究采用人表皮样癌细胞株、KB-31及其mdr 1转染细胞株、KB-G2和人甲状腺肿瘤细胞株、TCO-1，这些细胞株均过表达mdr 1。在Donald J Hnatowich博士（马萨诸塞州大学医学院教授）的监督下，用Tc-99 m标记反义DNA（AS）和正义DNA（S），然后将它们与KB-31、KB-G2和TCO-1细胞一起孵育。我们发现AS-积累是通过反义机制发生的.将Tc-99 m-AS注射到小鼠KB-G2肿瘤中，成功地获得了基因显像。这是世界上第一个反义显像的报道.静脉注射Tc-99 m-AS后，肿瘤特异性蓄积于KB-G2肿瘤内，但蓄积量不足以成像. JetPEI、Neophectin和Chariot等转染剂使Tc-99 m-AS在KB-G2细胞中的蓄积量增加了3-8倍，但在体内研究中没有观察到明显的增加.用我们独特的方法制备了由Tc-99 m-Streptavidine-TAT-AS组成的纳米颗粒，与裸AS-DNA相比，这些纳米颗粒显示出更高的细胞蓄积。目前正在进行体内研究。

项目成果

Initial Mechanistic Studies of Antisense Targeting in Cells

细胞反义靶向的初步机制研究

• 发表时间: 2006
• 期刊: Journal of Nuclear Medicine 47(2)
• 作者: Shigematsu N;Takami H;Kubo A;Xinrong Liu
• 通讯作者: Xinrong Liu

Evidence of antisense tumor targeting in mice

• DOI: 10.1021/bc0499073
• 发表时间: 2004-11-01
• 期刊: BIOCONJUGATE CHEMISTRY
• 影响因子: 4.7
• 作者: Nakamura, K;Fan, C;Hnatowich, DJ
• 通讯作者: Hnatowich, DJ

Improved Delivery in Cell Culture of Radiolabeled Antisense DNAs by Duplex Formation

• DOI: 10.1007/s11307-006-0050-7
• 发表时间: 2006-08
• 期刊: Molecular Imaging and Biology
• 影响因子: 3.1
• 作者: Xinrong Liu;Kayoko Nakamura;Yi Wang;Surong Zhang;Jiang He;Guozheng Liu;S. Dou;A. Kubo;M. Rusckowski;D. Hnatowich

Tumor Imaging in Mice with Tc-99m-Antisense DNA Administrated Intra-tumorally

瘤内注射 Tc-99m 反义 DNA 的小鼠肿瘤成像

• 发表时间: 2004
• 期刊: Journal of Nuclear Medicine 45, Suppl
• 作者: Nakamura K;et al.
• 通讯作者: et al.

遺伝子と臨床核医学 : 遺伝子組み換えとプラズミド

基因与临床核医学：基因重组和质粒

• 发表时间: 2004
• 期刊: 臨床核医学 37, 3
• 作者: E.Sato;E.Tanaka;H.Mori;T.Kawai;T.Inoue;A.Ogawa;M.Izumisawa;K.Takahashi;S.Sato;T.Ichimaru;K.Takayama;中村佳代子
• 通讯作者: 中村佳代子